DOCSLIB.ORG

Molecular Regulatory Mechanism of Exocytosis in the Salivary Glands

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Molecular Regulatory Mechanism of Exocytosis in the Salivary Glands International Journal of Molecular Sciences Review Molecular Regulatory Mechanism of Exocytosis in the Salivary Glands Akiko Suzuki 1,2 and Junichi Iwata 1,2,3,* 1 Department of Diagnostic & Biomedical Sciences, The University of Texas Health Science Center at Houston School of Dentistry, Houston, TX 77054, USA; [email protected] 2 Center for Craniofacial Research, The University of Texas Health Science Center at Houston School of Dentistry, Houston, TX 77054, USA 3 Program of Biochemistry and Cell Biology, The University of Texas Graduate School of Biomedical Sciences at Houston, Houston, TX 77030, USA * Correspondence: [email protected]; Tel.: +1-713-486-2641 Received: 1 September 2018; Accepted: 11 October 2018; Published: 17 October 2018 Abstract: Every day, salivary glands produce about 0.5 to 1.5 L of saliva, which contains salivary proteins that are essential for oral health. The contents of saliva, 0.3% proteins (1.5 to 4.5 g) in fluid, help prevent oral infections, provide lubrication, aid digestion, and maintain oral health. Acinar cells in the lobular salivary glands secrete prepackaged secretory granules that contain salivary components such as amylase, mucins, and immunoglobulins. Despite the important physiological functions of salivary proteins, we know very little about the regulatory mechanisms of their secretion via exocytosis, which is a process essential for the secretion of functional proteins, not only in salivary glands, but also in other secretory organs, including lacrimal and mammary glands, the pancreas, and prostate. In this review, we discuss recent findings that elucidate exocytosis by exocrine glands, especially focusing on the salivary glands, in physiological and pathological conditions. Keywords: exocytosis; salivary glands; Sjögren’s syndrome; protein secretion; membrane trafficking 1. Introduction The major (submandibular, sublingual, and parotid) and minor salivary glands produce 0.5–1.5 L of saliva daily, which is essential for oral health. Saliva comprises 99.5% of water, 0.3% of proteins, and 0.2% of both inorganic and organic substances [1]. The solid components differ from person to person, and from time to time in the same person. For example, salivary glands secrete 1.5 to 4.5 g proteins daily at a concentration of 0.47 ± 0.19 to 2.67 ± 0.54 mg/mL [2]. The protein concentration in saliva is lower than that in tears, which is around 8 mg/mL under basal conditions [3,4]. Acinar cells are a major cell type within the salivary glands, responsible for the production and secretion of prepackaged secretory granules that contain key functional salivary components, such as amylase, mucins, and immunoglobulins [5]. These salivary components are functionally important for the digestion and taste of foods, lubrication, buffering, and prevention of dental caries, periodontitis, candidiasis and halitosis (bad breath). This secretion process in exocrine glands, called exocytosis, involves secretory vesicle trafficking, docking, priming, and membrane fusion [6]. A failure during any of the steps in exocytosis in the salivary glands results in altered secretion of salivary proteins (Table1). Interestingly, several studies show that secretion of salivary proteins is reduced, and the contents of saliva are also changed, in patients with dry mouth syndromes (e.g., Sjögren’s syndrome) and in a mouse model for Sjögren’s syndrome (e.g., non-obese diabetic (NOD) mice) [1,7–10]. This evidence suggests that exocytosis may Int. J. Mol. Sci. 2018, 19, 3208; doi:10.3390/ijms19103208 www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2018, 19, 3208 2 of 16 be altered in various conditions, and the altered salivary contents may constitute a risk factor for major oral health issues [11,12]. There are two types of acinar cells—serous and mucous acinar—in the salivary glands. Serous acini are composed of typically 8–12 pyramid-shaped cells containing many secretory granules in the apical cytoplasm, a well-developed rough-endoplasmic reticulum (rough-ER), and pronounced round nuclei in the middle of the cytoplasm. They secrete serous saliva, which contains digestive α-amylase (AMY1A), a protein that is crucial for food digestion [13,14]. By contrast, mucous acinar cells are larger than serous acinar cells and have flat nuclei towards the basal cell surface due to large numbers of mucin granules aggregated in the apical cytoplasm [13,14]. Mucous saliva contains mucins that are important for lubrication and oral health [13,14]. The salivary glands secrete saliva rich in salivary proteins, the diverse functions of which maintain oral health by providing lubrication, initiating digestion, and offering first-line immunity. Therefore, disruption of salivary gland functions quickly results in widespread deterioration of oral health. Despite the important physiological functions of salivary proteins, we know very little about the regulatory mechanism(s) of exocytosis in the salivary glands. A detailed understanding of the mechanism(s) by which exocytosis is regulated will provide new knowledge of its key function(s), not only in the salivary glands but also in other secretory organs, including the lacrimal and mammary glands, the pancreas, and prostate, in physiological and pathological conditions. Ultimately, this approach will identify novel targets for therapeutics and contribute to new diagnostic tools for identifying exocytosis defects in at-risk populations such as those with high cholesterol levels or who have high-cholesterol diets. Sjögren’s syndrome is an autoimmune disorder characterized by lymphocytic infiltration of the exocrine glands, mainly the salivary and lacrimal glands, which results in reduced secretory functions and oral and ocular dryness [15,16]. The diagnosis is based on a combination of symptoms, physical examination, and blood tests (American-European Consensus Group, AECG, 2002; Sjögren’s International Collaborative Clinical Alliance, SICCA, 2012). While the pathogenesis of Sjögren’s syndrome remains elusive, various factors, including environmental, genetic and hormonal factors, seem to be involved, and either immune cells or exocrine gland cells are primarily damaged or disorganized to induce inflammation in the salivary and lacrimal glands [17–19]. Table 1. Phenotype in mice with deficiencies in genes related to the exocytosis process. Mutant Mouse Phenotype Gene Name Reference Salivary Acinar Cells Lacrimal Acinar Cells Pancreatic Acinar Cells Accumulation of secretory Accumulation of Accumulation of secretory Vamp8 [20] vesicles secretory vesicles vesicles Syntaxin2 Not studied Not studied Increased exocytosis [21] Enlarged secretory Rab3d Enlarged secretory vesicles vesicles Enlarged secretory vesicles [22,23] Decreased total protein amount in tears Increased total protein Rab27a Not studied Not studied [23,24] amount in tears Reduced number of Rab27b Not studied secretory vesicles Not studied [23,24] Increased total protein amount in tears Accumulation of enlarged Accumulation of enlarged Noc2 Not studied [25,26] secretory vesicles secretory vesicles Gland degeneration Gland degeneration Sec23b Not studied [27–29] Absent or reduced number Absent or reduced number of secretory vesicles of secretory vesicles Int. J. Mol. Sci. 2018, 19, x 3 of 16 Int. J. Mol. Sci. 2018, 19, 3208 3 of 16 2. The Exocytosis Process 2. TheExocytosis Exocytosis is Processa dynamic secretion process consisting of vesicle trafficking, tethering, docking, priming,Exocytosis and fusion is a [6,30]. dynamic After secretion proteins process are folded consisting in the ER, of they vesicle leave trafficking, this organelle tethering, inside docking, vesicles priming,coated with and the fusion coat [protein6,30]. Aftercomplex proteins II (COP areII) folded towards in the ER,Golgi they apparatus. leave this Through organelle the insidetrans- vesiclesGolgi network, coated withproteins the coatare proteintransported complex to variou II (COPII)s destinations towards theby Golgia variety apparatus. of mechanisms. Through For the transexample,-Golgi the network, destination proteins of the arediffe transportedrent proteins to is various determined destinations by the molecule by a variety coats ofon mechanisms. a vesicle, by Foractin-dependent example, the motors destination (Myosins), of the and different by microt proteinsubule-dependent is determined motors by (Kinesins the molecule and Dyneins) coats on a[31,32]. vesicle, During by actin-dependent intracellular transportation, motors (Myosins), RAB and GTPases by microtubule-dependent and their effectors regulate motors (Kinesinsthe secretory and Dyneins)vesicle movement [31,32]. During throughout intracellular the cytoskeleton transportation, [33]. RAB GTPases and their effectors regulate the secretorySoluble vesicle N-ethylmaleimide-sensitive movement throughout the factor cytoskeleton attachment [33]. protein (SNAP) receptor (SNARE) proteins,Soluble suchN -ethylmaleimide-sensitiveas vesicle-associated membrane factor attachmentproteins (VAMPs), protein (SNAP)are present receptor in the (SNARE) secretory proteins, vesicle suchmembrane as vesicle-associated (Figure 1). The membrane secretory vesicles proteins are (VAMPs), transported are present from the in the trans secretory-Golgi network vesicle membrane via actin- (Figureand microtubule-based1). The secretory molecular vesicles motor are transported proteins towards from the plasmatrans-Golgi membrane. network Once via the actin- secretory and microtubule-basedvesicles arrive to the molecular plasma motormembrane,
Load more

Recommended publications
  • Mea6 Controls VLDL Transport Through the Coordinated Regulation of COPII Assembly
    Cell Research (2016) 26:787-804. npg © 2016 IBCB, SIBS, CAS All rights reserved 1001-0602/16 $ 32.00 ORIGINAL ARTICLE www.nature.com/cr Mea6 controls VLDL transport through the coordinated regulation of COPII assembly Yaqing Wang1, *, Liang Liu1, 2, *, Hongsheng Zhang1, 2, Junwan Fan1, 2, Feng Zhang1, 2, Mei Yu3, Lei Shi1, Lin Yang1, Sin Man Lam1, Huimin Wang4, Xiaowei Chen4, Yingchun Wang1, Fei Gao5, Guanghou Shui1, Zhiheng Xu1, 6 1State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China; 2University of Chinese Academy of Sciences, Beijing 100101, China; 3School of Life Science, Shandong University, Jinan 250100, China; 4Institute of Molecular Medicine, Peking University, Beijing 100871, China; 5State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; 6Translation- al Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China Lipid accumulation, which may be caused by the disturbance in very low density lipoprotein (VLDL) secretion in the liver, can lead to fatty liver disease. VLDL is synthesized in endoplasmic reticulum (ER) and transported to Golgi apparatus for secretion into plasma. However, the underlying molecular mechanism for VLDL transport is still poor- ly understood. Here we show that hepatocyte-specific deletion of meningioma-expressed antigen 6 (Mea6)/cutaneous T cell lymphoma-associated antigen 5C (cTAGE5C) leads to severe fatty liver and hypolipemia in mice. Quantitative lip- idomic and proteomic analyses indicate that Mea6/cTAGE5 deletion impairs the secretion of different types of lipids and proteins, including VLDL, from the liver.
    [Show full text]
  • Identification of Secretory Granule Phosphatidylinositol 4,5-Bisphosphate- Interacting Proteins Using an Affinity Pulldown Strategy*□S
    Research Identification of Secretory Granule Phosphatidylinositol 4,5-Bisphosphate- interacting Proteins Using an Affinity Pulldown Strategy*□S Shona L. Osborne‡§, Tristan P. Wallis¶ʈ, Jose L. Jimenez**, Jeffrey J. Gorman¶ʈ, and Frederic A. Meunier‡§‡‡ Phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) syn- required for secretion is synthesized on the plasma mem- thesis is required for calcium-dependent exocytosis in neu- brane (4), and the binding of vesicle-associated proteins to rosecretory cells. We developed a PtdIns(4,5)P2 bead pull- PtdIns(4,5)P2 in trans is thought to be important for bringing down strategy combined with subcellular fractionation to the vesicle and plasma membranes together prior to exocy- Downloaded from identify endogenous chromaffin granule proteins that inter- tosis to ensure rapid and efficient fusion upon calcium influx act with PtdIns(4,5)P . We identified two synaptotagmin iso- 2 (5). Putative PtdIns(4,5)P effectors include synaptotagmin 1 forms, synaptotagmins 1 and 7; spectrin; ␣-adaptin; and 2 (Syt1) (6, 7), calcium-activated protein for secretion (CAPS) (8, synaptotagmin-like protein 4 (granuphilin) by mass spec- 9), and rabphilin (10). trometry and Western blotting. The interaction between https://www.mcponline.org CAPS binds to PtdIns(4,5)P2 and is a potential PtdIns(4,5)P2 synaptotagmin 7 and PtdIns(4,5)P2 and its functional rele- vance was investigated. The 45-kDa isoform of synaptotag- effector for priming of secretory granule exocytosis (9). How- min 7 was found to be highly expressed in adrenal chromaf- ever, recent work on the CAPS1 knock-out mouse highlighted fin cells compared with PC12 cells and to mainly localize to an involvement of CAPS1 in the refilling or storage of cate- secretory granules by subcellular fractionation, immunoiso- cholamine in mature secretory granules therefore suggesting lation, and immunocytochemistry.
    [Show full text]
  • Directed Proximity-Dependent Biotin Labelling in Zebrafish
    TOOLS AND RESOURCES In vivo proteomic mapping through GFP- directed proximity-dependent biotin labelling in zebrafish Zherui Xiong1, Harriet P Lo1, Kerrie-Ann McMahon1, Nick Martel1, Alun Jones1, Michelle M Hill2, Robert G Parton1,3*, Thomas E Hall1* 1Institute for Molecular Bioscience, The University of Queensland, Brisbane, Australia; 2QIMR Berghofer Medical Research Institute, Herston, Australia; 3Centre for Microscopy and Microanalysis, The University of Queensland, Brisbane, Australia Abstract Protein interaction networks are crucial for complex cellular processes. However, the elucidation of protein interactions occurring within highly specialised cells and tissues is challenging. Here, we describe the development, and application, of a new method for proximity- dependent biotin labelling in whole zebrafish. Using a conditionally stabilised GFP-binding nanobody to target a biotin ligase to GFP-labelled proteins of interest, we show tissue-specific proteomic profiling using existing GFP-tagged transgenic zebrafish lines. We demonstrate the applicability of this approach, termed BLITZ (Biotin Labelling In Tagged Zebrafish), in diverse cell types such as neurons and vascular endothelial cells. We applied this methodology to identify interactors of caveolar coat protein, cavins, in skeletal muscle. Using this system, we defined specific interaction networks within in vivo muscle cells for the closely related but functionally distinct Cavin4 and Cavin1 proteins. *For correspondence: Introduction [email protected] (RGP); [email protected] (TEH) The understanding of the biological functions of a protein requires detailed knowledge of the mole- cules with which it interacts. However, robust elucidation of interacting proteins, including not only Competing interests: The strong direct protein-protein interactions, but also weak, transient or indirect interactions is challeng- authors declare that no ing.
    [Show full text]
  • Synaptotagmin 7 Switches Short-Term Synaptic Plasticity from Depression
    www.nature.com/scientificreports OPEN Synaptotagmin 7 switches short‑term synaptic plasticity from depression to facilitation by suppressing synaptic transmission Takaaki Fujii1, Akira Sakurai1, J. Troy Littleton2 & Motojiro Yoshihara1* Short‑term synaptic plasticity is a fast and robust modifcation in neuronal presynaptic output that can enhance release strength to drive facilitation or diminish it to promote depression. The mechanisms that determine whether neurons display short‑term facilitation or depression are still unclear. Here we show that the Ca2+‑binding protein Synaptotagmin 7 (Syt7) determines the sign of short‑term synaptic plasticity by controlling the initial probability of synaptic vesicle (SV) fusion. Electrophysiological analysis of Syt7 null mutants at Drosophila embryonic neuromuscular junctions demonstrate loss of the protein converts the normally observed synaptic facilitation response during repetitive stimulation into synaptic depression. In contrast, overexpression of Syt7 dramatically enhanced the magnitude of short‑term facilitation. These changes in short‑term plasticity were mirrored by corresponding alterations in the initial evoked response, with SV release probability enhanced in Syt7 mutants and suppressed following Syt7 overexpression. Indeed, Syt7 mutants were able to display facilitation in lower [Ca2+] where release was reduced. These data suggest Syt7 does not act by directly sensing residual Ca2+ and argues for the existence of a distinct Ca2+ sensor beyond Syt7 that mediates facilitation. Instead, Syt7 normally suppresses synaptic transmission to maintain an output range where facilitation is available to the neuron. Synaptotagmins (Syts) are a large family of Ca2+ binding proteins, with the Syt1 isoform functioning as the major Ca2+ sensor for synchronous synaptic vesicle (SV) fusion1. Ca2+ also controls presynaptic forms of short-term plasticity, with other Syt isoforms representing promising candidates to mediate these processes.
    [Show full text]
  • Communication Between the Endoplasmic Reticulum and Peroxisomes in Mammalian Cells
    Communication between the Endoplasmic Reticulum and Peroxisomes in Mammalian Cells by Rong Hua A thesis submitted in conformity with the requirements for the degree of Doctor of Philosophy Department of Biochemistry University of Toronto © Copyright by Rong Hua (2017) ii Communication between the Endoplasmic Reticulum and Peroxisomes in Mammalian Cells Rong Hua Doctor of Philosophy Department of Biochemistry University of Toronto 2017 Abstract Peroxisomes are important metabolic organelles found in virtually all eukaryotic cells. Since their discovery, peroxisomes have long been seen in close proximity to the endoplasmic reticulum (ER). The interplay between the two organelles is suggested to be important for peroxisome biogenesis as the ER may serve as a source for both lipids and peroxisomal membrane proteins (PMPs) for peroxisome growth and maintenance. On the other hand, various lipid molecules are exchanged between them for the biosynthesis of specialized lipids such as bile acids, plasmalogens and cholesterol. However, how proteins and lipids are transported between the two organelles is not yet fully understood. Previously, the peroxisomal biogenesis factor PEX16 was shown to serve as a receptor for PMPs in the ER and also as a mediator of the subsequent transport of these ER-targeted PMPs to peroxisomes. Here, I extended these results by carrying out a comprehensive mutational analysis of PEX16 aimed at gaining insights into the molecular targeting signals responsible for its ER-to-peroxisome trafficking and the domain(s) involved in its PMP recruitment function at the ER. I also showed that the recruitment function of PEX16 is conserved in plants. To gain further mechanistic insight into PEX16 function, the proteins proximal to iii PEX16 were identified using the proximity-dependent BioID analysis.
    [Show full text]
  • A Minimum-Labeling Approach for Reconstructing Protein Networks Across Multiple Conditions
    A Minimum-Labeling Approach for Reconstructing Protein Networks across Multiple Conditions Arnon Mazza1, Irit Gat-Viks2, Hesso Farhan3, and Roded Sharan1 1 Blavatnik School of Computer Science, Tel Aviv University, Tel Aviv 69978, Israel. Email: [email protected]. 2 Dept. of Cell Research and Immunology, Tel Aviv University, Tel Aviv 69978, Israel. 3 Biotechnology Institute Thurgau, University of Konstanz, Unterseestrasse 47, CH-8280 Kreuzlingen, Switzerland. Abstract. The sheer amounts of biological data that are generated in recent years have driven the development of network analysis tools to fa- cilitate the interpretation and representation of these data. A fundamen- tal challenge in this domain is the reconstruction of a protein-protein sub- network that underlies a process of interest from a genome-wide screen of associated genes. Despite intense work in this area, current algorith- mic approaches are largely limited to analyzing a single screen and are, thus, unable to account for information on condition-specific genes, or reveal the dynamics (over time or condition) of the process in question. Here we propose a novel formulation for network reconstruction from multiple-condition data and devise an efficient integer program solution for it. We apply our algorithm to analyze the response to influenza in- fection in humans over time as well as to analyze a pair of ER export related screens in humans. By comparing to an extant, single-condition tool we demonstrate the power of our new approach in integrating data from multiple conditions in a compact and coherent manner, capturing the dynamics of the underlying processes. 1 Introduction With the increasing availability of high-throughput data, network biol- arXiv:1307.7803v1 [q-bio.QM] 30 Jul 2013 ogy has become the method of choice for filtering, interpreting and rep- resenting these data.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Supplementary Materials
    1 Supplementary Materials: Supplemental Figure 1. Gene expression profiles of kidneys in the Fcgr2b-/- and Fcgr2b-/-. Stinggt/gt mice. (A) A heat map of microarray data show the genes that significantly changed up to 2 fold compared between Fcgr2b-/- and Fcgr2b-/-. Stinggt/gt mice (N=4 mice per group; p<0.05). Data show in log2 (sample/wild-type). 2 Supplemental Figure 2. Sting signaling is essential for immuno-phenotypes of the Fcgr2b-/-lupus mice. (A-C) Flow cytometry analysis of splenocytes isolated from wild-type, Fcgr2b-/- and Fcgr2b-/-. Stinggt/gt mice at the age of 6-7 months (N= 13-14 per group). Data shown in the percentage of (A) CD4+ ICOS+ cells, (B) B220+ I-Ab+ cells and (C) CD138+ cells. Data show as mean ± SEM (*p < 0.05, **p<0.01 and ***p<0.001). 3 Supplemental Figure 3. Phenotypes of Sting activated dendritic cells. (A) Representative of western blot analysis from immunoprecipitation with Sting of Fcgr2b-/- mice (N= 4). The band was shown in STING protein of activated BMDC with DMXAA at 0, 3 and 6 hr. and phosphorylation of STING at Ser357. (B) Mass spectra of phosphorylation of STING at Ser357 of activated BMDC from Fcgr2b-/- mice after stimulated with DMXAA for 3 hour and followed by immunoprecipitation with STING. (C) Sting-activated BMDC were co-cultured with LYN inhibitor PP2 and analyzed by flow cytometry, which showed the mean fluorescence intensity (MFI) of IAb expressing DC (N = 3 mice per group). 4 Supplemental Table 1. Lists of up and down of regulated proteins Accession No.
    [Show full text]
  • Protein Identities in Evs Isolated from U87-MG GBM Cells As Determined by NG LC-MS/MS
    Protein identities in EVs isolated from U87-MG GBM cells as determined by NG LC-MS/MS. No. Accession Description Σ Coverage Σ# Proteins Σ# Unique Peptides Σ# Peptides Σ# PSMs # AAs MW [kDa] calc. pI 1 A8MS94 Putative golgin subfamily A member 2-like protein 5 OS=Homo sapiens PE=5 SV=2 - [GG2L5_HUMAN] 100 1 1 7 88 110 12,03704523 5,681152344 2 P60660 Myosin light polypeptide 6 OS=Homo sapiens GN=MYL6 PE=1 SV=2 - [MYL6_HUMAN] 100 3 5 17 173 151 16,91913397 4,652832031 3 Q6ZYL4 General transcription factor IIH subunit 5 OS=Homo sapiens GN=GTF2H5 PE=1 SV=1 - [TF2H5_HUMAN] 98,59 1 1 4 13 71 8,048185945 4,652832031 4 P60709 Actin, cytoplasmic 1 OS=Homo sapiens GN=ACTB PE=1 SV=1 - [ACTB_HUMAN] 97,6 5 5 35 917 375 41,70973209 5,478027344 5 P13489 Ribonuclease inhibitor OS=Homo sapiens GN=RNH1 PE=1 SV=2 - [RINI_HUMAN] 96,75 1 12 37 173 461 49,94108966 4,817871094 6 P09382 Galectin-1 OS=Homo sapiens GN=LGALS1 PE=1 SV=2 - [LEG1_HUMAN] 96,3 1 7 14 283 135 14,70620005 5,503417969 7 P60174 Triosephosphate isomerase OS=Homo sapiens GN=TPI1 PE=1 SV=3 - [TPIS_HUMAN] 95,1 3 16 25 375 286 30,77169764 5,922363281 8 P04406 Glyceraldehyde-3-phosphate dehydrogenase OS=Homo sapiens GN=GAPDH PE=1 SV=3 - [G3P_HUMAN] 94,63 2 13 31 509 335 36,03039959 8,455566406 9 Q15185 Prostaglandin E synthase 3 OS=Homo sapiens GN=PTGES3 PE=1 SV=1 - [TEBP_HUMAN] 93,13 1 5 12 74 160 18,68541938 4,538574219 10 P09417 Dihydropteridine reductase OS=Homo sapiens GN=QDPR PE=1 SV=2 - [DHPR_HUMAN] 93,03 1 1 17 69 244 25,77302971 7,371582031 11 P01911 HLA class II histocompatibility antigen,
    [Show full text]
  • RNF11 at the Crossroads of Protein Ubiquitination
    biomolecules Review RNF11 at the Crossroads of Protein Ubiquitination Anna Mattioni, Luisa Castagnoli and Elena Santonico * Department of Biology, University of Rome Tor Vergata, Via della ricerca scientifica, 00133 Rome, Italy; [email protected] (A.M.); [email protected] (L.C.) * Correspondence: [email protected] Received: 29 September 2020; Accepted: 8 November 2020; Published: 11 November 2020 Abstract: RNF11 (Ring Finger Protein 11) is a 154 amino-acid long protein that contains a RING-H2 domain, whose sequence has remained substantially unchanged throughout vertebrate evolution. RNF11 has drawn attention as a modulator of protein degradation by HECT E3 ligases. Indeed, the large number of substrates that are regulated by HECT ligases, such as ITCH, SMURF1/2, WWP1/2, and NEDD4, and their role in turning off the signaling by ubiquitin-mediated degradation, candidates RNF11 as the master regulator of a plethora of signaling pathways. Starting from the analysis of the primary sequence motifs and from the list of RNF11 protein partners, we summarize the evidence implicating RNF11 as an important player in modulating ubiquitin-regulated processes that are involved in transforming growth factor beta (TGF-β), nuclear factor-κB (NF-κB), and Epidermal Growth Factor (EGF) signaling pathways. This connection appears to be particularly significant, since RNF11 is overexpressed in several tumors, even though its role as tumor growth inhibitor or promoter is still controversial. The review highlights the different facets and peculiarities of this unconventional small RING-E3 ligase and its implication in tumorigenesis, invasion, neuroinflammation, and cancer metastasis. Keywords: Ring Finger Protein 11; HECT ligases; ubiquitination 1.
    [Show full text]
  • Supplementary Figures 1-14 and Supplementary References
    SUPPORTING INFORMATION Spatial Cross-Talk Between Oxidative Stress and DNA Replication in Human Fibroblasts Marko Radulovic,1,2 Noor O Baqader,1 Kai Stoeber,3† and Jasminka Godovac-Zimmermann1* 1Division of Medicine, University College London, Center for Nephrology, Royal Free Campus, Rowland Hill Street, London, NW3 2PF, UK. 2Insitute of Oncology and Radiology, Pasterova 14, 11000 Belgrade, Serbia 3Research Department of Pathology and UCL Cancer Institute, Rockefeller Building, University College London, University Street, London WC1E 6JJ, UK †Present Address: Shionogi Europe, 33 Kingsway, Holborn, London WC2B 6UF, UK TABLE OF CONTENTS 1. Supplementary Figures 1-14 and Supplementary References. Figure S-1. Network and joint spatial razor plot for 18 enzymes of glycolysis and the pentose phosphate shunt. Figure S-2. Correlation of SILAC ratios between OXS and OAC for proteins assigned to the SAME class. Figure S-3. Overlap matrix (r = 1) for groups of CORUM complexes containing 19 proteins of the 49-set. Figure S-4. Joint spatial razor plots for the Nop56p complex and FIB-associated complex involved in ribosome biogenesis. Figure S-5. Analysis of the response of emerin nuclear envelope complexes to OXS and OAC. Figure S-6. Joint spatial razor plots for the CCT protein folding complex, ATP synthase and V-Type ATPase. Figure S-7. Joint spatial razor plots showing changes in subcellular abundance and compartmental distribution for proteins annotated by GO to nucleocytoplasmic transport (GO:0006913). Figure S-8. Joint spatial razor plots showing changes in subcellular abundance and compartmental distribution for proteins annotated to endocytosis (GO:0006897). Figure S-9. Joint spatial razor plots for 401-set proteins annotated by GO to small GTPase mediated signal transduction (GO:0007264) and/or GTPase activity (GO:0003924).
    [Show full text]
  • Uncoupling the Functions of CALM in VAMP Sorting and Clathrin-Coated Pit Formation
    Uncoupling the Functions of CALM in VAMP Sorting and Clathrin-Coated Pit Formation Daniela A. Sahlender¤a, Patrycja Kozik¤b, Sharon E. Miller, Andrew A. Peden¤c, Margaret S. Robinson* Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom Abstract CALM (clathrin assembly lymphoid myeloid leukemia protein) is a cargo-selective adaptor for the post-Golgi R-SNAREs VAMPs 2, 3, and 8, and it also regulates the size of clathrin-coated pits and vesicles at the plasma membrane. The present study has two objectives: to determine whether CALM can sort additional VAMPs, and to investigate whether VAMP sorting contributes to CALM-dependent vesicle size regulation. Using a flow cytometry-based endocytosis efficiency assay, we demonstrate that CALM is also able to sort VAMPs 4 and 7, even though they have sorting signals for other clathrin adaptors. CALM homologues are present in nearly every eukaryote, suggesting that the CALM family may have evolved as adaptors for retrieving all post-Golgi VAMPs from the plasma membrane. Using a knockdown/rescue system, we show that wild-type CALM restores normal VAMP sorting in CALM-depleted cells, but that two non-VAMP-binding mutants do not. However, when we assayed the effect of CALM depletion on coated pit morphology, using a fluorescence microscopy- based assay, we found that the two mutants were as effective as wild-type CALM. Thus, we can uncouple the sorting function of CALM from its structural role. Citation: Sahlender DA, Kozik P, Miller SE, Peden AA, Robinson MS (2013) Uncoupling the Functions of CALM in VAMP Sorting and Clathrin-Coated Pit Formation.
    [Show full text]